国内首见三株棘状外瓶霉的研究

作者研究了国内首见的三株棘状外瓶霉[Exophiala spinifera(Nielsen et Conant)McGinms],观察了菌落形态,研究了此菌在光学显微镜下和扫描电镜下的特点,并作了外抗原试验。在光学显微镜下,本菌有暗色长棘状的分生孢子梗。在扫描电镜下,有很长的环痕梗,其尖端的环痕数目可达30个以上。有的环痕梗和甄氏外瓶霉[Exophiala jeanselmei(Langeron)McGinnis et Padhye]的环痕梗很相似。有些酵母样细胞上也可产生短的环痕产孢尖端。有一株菌的产孢细胞顶端有几个突起的环痕产孢尖端,呈假单轴性排列。另一株菌产生瓶梗。因此考虑此菌是一种多形性真菌。外抗原试验,一株菌符合棘状外瓶霉,另二株菌符合外瓶霉。     

长白山分解纤维素的瓶霉属和外瓶霉属真菌

瓶霉属、外瓶霉属真菌在自然界中广泛分布,是有着重要经济意义的一类真菌。迄今,我国已报道疣状瓶霉（Ｐｈｉａｌｏｐｈｏｒａ ｖｅｒｒｕｃｏｓａ）。裴氏瓶霉（Ｐｈ．ｐｅｄｒｏｓｏｉ）,棘状外瓶霉（Ｅｘｏｐｈｉａｌａ ｓｐｉｎｉｆｅｒａ）、皮炎外瓶霉（Ｅ．ｄｅｒｍａｔｉｔｉｄｉｓ）和甄氏外瓶霉（Ｅ．ｊｅａｎｓｅｌｍｅｉ＝Ｐｈ．ｇｏｕｇｅｒｏｔｉｉ）５种,均是分离自人体的病原真菌。在长白山自然保护区的原始林     

首报Phialophora expanda致着色芽生菌病伴CARD9缺陷1例

正着色芽生菌病(chromoblastomycosis)是一种由暗色真菌引起的累及皮肤和皮下组织的慢性肉芽肿性疾病。在我国北方其致病菌主要是卡氏枝孢瓶霉(Cladophialophora carrionii),南方主要是单梗着色霉(Fonsecaea monophora)和裴氏着色霉(Fonsecaea pedrosoi)~[1-2]。国内外目前尚未有CARD9缺陷致着色芽生菌病的报道;此外,国内尚无扩展瓶霉     

长白山分解纤维素的瓶霉属和外瓶霉属真菌

瓶霉属、外瓶霉属真菌在自然界中广泛分布,是有着重要经济意义的一类真菌。迄今,我国已报道疣状瓶霉（Phialophoraverrucosa）、裴氏瓶霉（Ph.pedrosoi）、棘状外瓶霉（Exophialaspinifera）、皮炎外瓶霉（E.dermatitidis）和甄氏外瓶霉（E.jeanselmei＝Ph．gougerotii）5种,均是分离自人体的病原真菌。在长白山自然保护区的原始林中,用生长锥取样器随机钻取腐朽林木髓心,在实验室进行分离培养。共鉴定出瓶霉属真菌2种,外瓶霉属真菌3种,其中美州瓶霉（Ph.americana、烂木瓶霉（Ph．richardsiae）和鲑外瓶霉（E.salmonis）为国内新记录种。     

棘状外瓶霉核糖体基因的初步研究

外瓶霉可致人类感染 ,不同生物群落的菌种 ,其致病性、药敏性等特征具有差异性。通过对 1 0株棘状外瓶霉核糖体基因及其转录间隔区进行序列测定 ,并与GeneBank中 9株同种真菌对比分析 ,揭示了不同生物群落的棘状外瓶霉虽然形态学差异性小 ,但在基因学上具有差异性 ;原属于甄氏外瓶霉变种的BMU 0 0 0 45 7(ATCC 2 41 5 2E .jeanselmeivar.hetoromorpha)与 2株棘状外瓶霉具有 1 0 0 %的同源性。研究提示了形态学特征相似的棘状外瓶霉在基因水平上具有差异性 ,核糖体基因及其转录间隔区对于研究菌群特性具有一定意义。     

皮炎外瓶霉导致的真菌性腹膜炎1例

报道1例由皮炎外瓶霉(Exophiala dermatitidis,Ed)导致的真菌性腹膜炎。此菌分离于1名长期腹膜透析患者的腹水。皮炎外瓶霉主要引起皮肤及皮下组织、中枢神经系统感染,引起腹膜炎者少见。现对此临床分离株进行真菌学分析、鉴定、药敏实验。患者经拔除透析管,改血液透析,同时抗真菌治疗,口服伊曲康唑0.2g/d,症状明显改善。     

临床皮炎外瓶霉荧光定量PCR检测方法的建立

目的 建立基于TaqMan探针技术的皮炎外瓶霉荧光定量PCR检测方法.方法 通过对皮炎外瓶霉ITS区域基因组序列（GenBank：JN675373.1）进行分析,设计合成特异性引物和荧光标记探针,优化荧光定量PCR反应条件.以临床标本中分离的皮炎外瓶霉为阳性菌株,及其他种类真菌和细菌作为阴性对照菌株,从特异性、敏感性及重复性方面对该方法检测效果进行评价.结果 该研究设计的引物和探针能扩增皮炎外瓶霉特异性序列.临床分离得到的皮炎外瓶霉在反应中有明显扩增曲线,而甄氏外瓶霉、棘状外瓶霉、烟曲霉、白色念珠菌、新生隐球菌、马内菲青霉等20株菌在CT值≤38范围内均未有扩增;利用基因重组构建的标准品完成了标准曲线的绘制,在1.0×103～1.0×107拷贝数（Cp）内具有良好的线性关系（R2=1.000）,最低可检出量为10 Cp/μL.结论 成功建立了荧光定量PCR检测皮炎外瓶霉方法,该法特异度强、敏感度高、重复性好,将有助于临床皮炎外瓶霉感染的早期诊断和针对性治疗.     

裴氏着色霉的扫描电镜研究

用扫描电镜研究了引起着色芽生菌病(chromoblastomycosis)的致病菌之一——裴氏着色霉(Fonsecaea pedrosoi)孢子的个体发生,除一级产孢细胞外,还有二级产孢细胞,喙枝孢型的分生孢子梗也可发育成为喙枝孢型的二级产孢细胞。     

全球皮炎外瓶霉感染流行现状的回顾性分析

目的分析皮炎外瓶霉感染的流行病学和临床特征,为提高皮炎外瓶霉感染的诊治水平提供科学依据。方法采用文献回顾和荟萃分析的方法,分析全球范围内已报道的皮炎外瓶霉感染病例的国籍、性别、年龄分布、危险因素、发病部位、临床表现、诊疗方法及预后等流行病学和临床特征。结果皮炎外瓶霉感染在免疫功能正常和免疫功能缺陷患者中均可发生,患者的男女性别比为1.10∶1.00,最常发病年龄段为51~60岁,肺(27.90%,17/61)为最常受累的器官,但不同地域的病例感染器官存在差异。约半数(47.54%,29/61)病例伴有各种免疫抑制的基础疾病或危险因素。皮炎外瓶霉感染的临床确诊主要依赖培养和分子鉴定,系统感染患者推荐联合伊曲康唑和特比萘芬作为抗真菌治疗方案。结论近年来皮炎外瓶霉感染的发病率在全球范围内呈上升趋势,肺部为系统感染患者最常受累的器官,诊断主要依赖于真菌培养。加强皮炎外瓶霉菌株的药敏监测和分子流行病学研究,对于提高皮炎外瓶霉感染的临床诊治水平非常必要。     

外瓶霉属真菌耐药性和降解能力的研究

外瓶霉属(Exophiala)真菌对五氯酚和杂酚油都有很强的耐药性,其中以菌株P-3182对五氯酚和菌株P-2830、P-3182对杂酚油的耐药性最强。外瓶霉属真菌不但能在五氯酚和杂酚油溶液中生长,而且具有降解这两种防腐剂的能力。而在砷铬酸铜溶液中不能生长,对环烷酸铜和砷铬酸铜的耐药性与对照菌种相似或稍高。     

Identification of pathogenic dematiaceous fungi and related taxa based on large subunit ribosomal DNA D1/D2 domain sequence analysis

The nucleotide sequences of the D1/D2 domains of large subunit (26S) ribosomal DNA for 76 strains of 46 species of pathogenic dematiaceous fungi and related taxa were determined. Intra-species sequence diversity of medically important dematiaceous fungi including Phialophora verrucosa, Fonsecaea pedrosoi, Fonsecaea compacta, Cladophialophora carrionii, Cladophialophora bantiana, Exophiala dermatitidis, Exophiala jeanselmei, Exophiala spinifera, Exophiala moniliae, and Hortaea werneckii were extremely small; as few as 0 changes were detected in C. bantiana, Fonsecaea and Exophiala species, 1 bp in C. carrionii and H. werneckii, and 2 bp in P. verrucosa. Inter-species nucleotide diversity between most species was higher. These data suggested that the D1/D2 domain is sufficiently variable for identification of pathogenic dematiaceous fungi and relevant species. The phylogenetic trees constructed from the sequence data revealed that most human pathogenic species formed a single cluster and that Cladosporium and Phialophora species were distributed polyphyletically into several clusters.     

Evaluation of skin test for chromoblastomycosis using antigens prepared from culture filtrates ofFonsecaea pedrosoi,Phialophora verrucosa,Wangiella dermatitidis andExophiala jeanselmei

Antigenic substances were prepared from culture filtrates ofFonsecaea pedrosoi, Phialophora verrucosa, Wangiella dermatitidis andExophiala jeanselmei. These antigenic substances were evaluated for detecting cutaneous delayed hypersensitivity in rats experimentally-infected withF. pedrosoi, P. verrucosa, W. dermatitidis, E. jeanselmei, Cladosporium carrionii andFonsecaea compactum and in patients with chromoblastomycosis caused byF. pedrosoi. TheF. pedrosoi antigen elicited positive reactions in all of the animals infected withF. pedrosoi and in 5 of 6 patients. TheP. verrucosa, W. dermatitidis andE. jeanselmei antigens elicited positive reactions in all of the animals infected with the homologous species. These antigens displayed cross-reactivity in some of the animals and patients, whereas more than half of them exhibited positive reactions only to the antigens prepared from the homologous species. These results suggest that a delayed-type skin test using the antigens prepared by the authors may be useful not only for the diagnosis of chromoblastomycosis but also for the identification of species of the causative agent.     

Morphologic and physiologic characteristics of Costa Rica pathogenic fungi (Dermatiaceae)]

Forty one strains of dematiaceous fungi from the Mycology collection of the University of Costa Rica were studied. Thirty three were pathogenic (Fonsecaea pedrosoi, Cladosporium carrionii, Xylohypha bantiana, Exophiala jeanselmei, Rhinocladiella aquaspersa, Phialophora verrucosa) and the other eight were contaminants (Hormodendrum sp.). Morphological studies were done using the slide culture technique. The physiological criteria used were: urease production, gelatin and Loeffler media liquefaction; xanthine, tyrosine, starch and casein hydrolysis; nitrate utilization; carbohydrate uptake; sensitivity to cycloheximide and thermotolerance in glucose-Sabouraud medium. The physiological tests did not provide characteristic patterns for the different genera of pathogenic fungi, even though these differences were detected in non pathogenic fungi; the tests may be useful for the quick separation of both groups. Physiological test may have a limited value in the identification of fungi and the morphological analysis cannot be substituted by physiological studies.     

Chromoblastomycosis – A clinical and mycological study of 71 cases from Sri Lanka

Chromoblastomycosis, a well-documented chronic fungal infection, represents a specific clinical entity with typical warty cutaneous nodules and a worldwide distribution. Although more prevalent in tropical and subtropical regions, only a few reports are available from Sri Lanka or from Asia. Five etiologic agents of chromoblastomycosis have been recognized worldwide. Of these the majority of infections have been caused by Fonsecaea pedrosoi. During the period from 1952 to 1962, only twelve culturally proven cases of this disease had been recorded from Sri Lanka. The fungus responsible was F. pedrosoi. The present report presents a study of the clinical and mycological features of 71 Sri Lankan patients with chromoblastomycosis for the 16-year period from 1978 to 1993. It documents three etiological agents. Culture identification was made in 69 cases. The three fungal species were Fonsecaea pedrosoi (64), Phialophora verrucosa (3) and a fungus compatible morphologically with F. compacta (2). The isolation of a fungus morphologically compatible with F. compacta is of significance since only 12 cases have been documented in the world's literature so far. This revised version was published online in June 2006 with corrections to the Cover Date.     

Mitochondrial DNA analysis of Exophiala jeanselmei and Exophiala dermatitidis

Mitochondrial DNA (mtDNA) analysis with Hae III, Hind. III and Msp I was performed in 45 Exophiala jeanselmei strains (30 Phialophora jeanselmei and 15 Phialophora gougerotii strains) and 31 Exophiala dermatitidis strains. The results were as follows, 1) P. jeanselmei and P. gougerotii are identical, 2) E. jeanselmei is classified into 18 types based on restriction profiles, 3) two strains of E. jeanselmei CBS 577.76 and CBS 578.76 are identified as E. dermatitidis, 4) E. dermatitidis has no intraspecific variation and is definitely distinct from E. jeanselmei, 5) E. jeanselmei is suggested to be a complex organism because of extensive mtDNA polymorphism.     

Granulomatous reactions induced by lipids extracted from Fonsecaea pedrosoi, Fonsecaea compactum, Cladosporium carrionii and Phialophora verrucosum

Granulomatous reactions induced by lipid extracts from the dermatophyte fungi Fonsecaea pedrosoi, Fonsecaea compactum, Cladosporium carrionii and Phialophora verrucosum, the causal organisms of chromoblastomycosis, were studied. Charcoal particles coated with the lipid extracts were prepared and injected intravenously into mice. Inflammation was characterized by an intense mononuclear cell infiltrate that lodged in the lung from 4 to 8 d after inoculation.     

Nutritional physiology of type isolates of currently accepted species ofExophiala andPhaeococcomyces

Nutritional physiological and tolerance tests were performed for all type strains of species currently classified in the black yeast generaExophiala andPhaeococcomyces, including some additional type strains of taxa recently reidentified asExophiala species. Most describedExophiala species can be distinguished by physiological characters.Exophiala jeanselmei with its varieties, andE. castellanii should all be retained as separate taxa. The pairs of strainsMycotorula schawii/Exophiala dermatitidis, Hormodendrum negronii/Exophiala jeanselmei var.lecaniicorni andSporotrichum gougerotii/Torula bergeri were found to be conspecific. Phenetic analyses of physiological data support the identity ofPhaeococcomyces exophialae as a yeast-like synanamorph ofExophiala spinifera. The taxonomic positions of the generaNadsoniella, Phaeoannellomyces andWangiella are discussed. The generaExophiala andPhaeococcomyces are unrelated.     

Taxonomy and morphology of dematiaceous fungi isolated from nature

The Hughes system of classification was applied to the identification of dematiaceous fungi isolated from nature. The majority of the isolates encountered could be identified with this system using brightfield microscopy. However, phase contrast microscopy was needed to determine the mode of conidial ontogeny with some species of Phialophora, Exophiala and Wangiella. The identifications based upon phase contrast microscopy were confirmed using the scanning electron microscope (SEM). Scanning electron photomicrographs are presented for Bispora betulina, Cladosporium trichoides, Exophiala jeanselmei, Phialophora revens, P. verrucosa and Wangiella dermatitidis. SEM observations of W. dermatitidis support the present classification of this organism as proposed by other workers. It is suggested that the Hughes system of classification and phase contrast microscopy be used to facilitate identification of isolates of dematiaceous fungi encountered in the clinical laboratory.